Protease Activated Receptor Signaling Is Required for African Trypanosome Traversal of Human Brain Microvascular Endothelial Cells 英文参考文献.docVIP

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Protease Activated Receptor Signaling Is Required for African Trypanosome Traversal of Human Brain Microvascular Endothelial Cells 英文参考文献.doc

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Protease Activated Receptor Signaling Is Required for African Trypanosome Traversal of Human Brain Microvascular Endothelial Cells 英文参考文献

ProteaseActivatedReceptorSignalingIsRequiredfor AfricanTrypanosomeTraversalofHumanBrain MicrovascularEndothelialCells DennisJ.Grab1,2*,JoseC.Garcia-Garcia2,OlgaV.Nikolskaia1,YuriV.Kim1¤,AmandaBrown3,CarlosA. Pardo3,YongqingZhang4,KevinG.Becker4,BrendaA.Wilson5,AnaPaulaC.deA.Lima6 ,Julio Scharfstein6,J.StephenDumler2 1Department of Pediatrics, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 2Department of Pathology, The Johns HopkinsUniversitySchoolofMedicine,Baltimore,Maryland,United StatesofAmerica, 3DepartmentofNeurology,TheJohnsHopkinsUniversitySchoolofMedicine, Baltimore, Maryland, United States of America, 4Gene Expression and Genomics Unit, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States ofAmerica, 5Department ofMicrobiology, University ofIllinois atUrbana-Champaign, Urbana, Illinois, United States ofAmerica, 6Instituto deBiof?′sica CarlosChagasFilho,UniversidadeFederaldoRiodeJaneiro,RiodeJaneiro,Brazil Abstract Background: Using human brain microvascular endothelial cells (HBMECs) as an in vitro model for how African trypanosomes cross the human blood-brain barrier (BBB) we recently reported that the parasites cross the BBB by generatingcalciumactivationsignalsinHBMECsthroughtheactivityofparasitecysteineproteases,particularlycathepsinL (brucipain).Inthecurrentstudy,weexaminedthepossibleroleofaclassofproteasestimulatedHBMECGproteincoupled receptors(GPCRs)knownasproteaseactivatedreceptors(PARs)thatmightbeimplicatedincalciumsignalingbyAfrican trypanosomes. Methodology/PrincipalFindings:UsingRNAinterference(RNAi)wefoundthatinvitroPAR-2gene(F2RL1)expressionin HBMECmonolayerscould bereducedbyover95%.Wealsofoundthattheability of Trypanosomabruceirhodesiense to crossF2RL1-silencedHBMECmonolayerswasreduced(39%–49%)andthatHBMECssilencedforF2RL1maintainedcontrol levelsofbarrierfunctioninthepresenceoftheparasite.ConsistentwiththeroleofPAR-2,wefoundthatHBMECbarrier